Previous title: Inhibition of ex vivo VEGF-induced angiogenesis by tyrosol and hydroxytyrosol present in Mediterranean diet foods: Quantitative three-dimensional mouse aortic ring model Current version title: Inhibition of ex vivo VEGF-induced angiogenesis by tyrosol and hydroxytyrosol: Quantitative three-dimensional mouse aortic ring mode

Abstract

Mediterranean diet foods such as olives, virgin olive oil and wine are sources of tyrosol (TOL) and hydroxytyrosol (HT) bioactive compounds. HT has already shown in vitro antiangiogenic effects in HUVEC cells. Since TOL is structurally closely related to HT, the aim of the present study was firstly to evaluate the anti-angiogenic properties of TOL regarding inhibition of VEGF-induced VEGFR2 phosphorylation as well as its effects on intracellular signaling cascade (PLCγ1, Akt and eNOS). Additionally, this paper aims to demonstrate the anti-angiogenic effects of HT and TOL using the ex vivo gold standard mouse aortic rings model. Our results have demonstrated that TOL significantly inhibit VEGF-induced VEGFR2 activation in HUVEC cells, with an IC50 value of 38.33 µM. Additionally, TOL completely blocked PLCγ1 activation, a key component of the VEGFR-2-mediated signalling pathway, while simultaneously increased the phosphorylation of Akt and eNOS, critical molecules in the regulation of angiogenesis and vasodilation. This study is the first to use the mouse aortic ring model to demonstrate the anti-angiogenic effect of TOL and HT. A significant reduction of capillary sprouting at 68% and 96% was observed for TOL and HT, respectively. These results not only support the potential of TOL and HT as natural antiangiogenic agents but also offer a new perspective on how diet, especially Mediterranean diet may influence the prevention and treatment of angiogenesis-related diseases, such as cancer and cardiovascular diseases.